CA2477433A1 - Methods and materials for detecting genetic material - Google Patents
Methods and materials for detecting genetic material Download PDFInfo
- Publication number
- CA2477433A1 CA2477433A1 CA002477433A CA2477433A CA2477433A1 CA 2477433 A1 CA2477433 A1 CA 2477433A1 CA 002477433 A CA002477433 A CA 002477433A CA 2477433 A CA2477433 A CA 2477433A CA 2477433 A1 CA2477433 A1 CA 2477433A1
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- detergent
- layer
- edta
- isolation layer
- weak base
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1017—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by filtration, e.g. using filters, frits, membranes
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- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Crystallography & Structural Chemistry (AREA)
- Plant Pathology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
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- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
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Abstract
In accordance with the present invention, there is provided a device, method and kit for detecting any genetic material containing moieties. The present invention enables identification of known or unknown virions or bacteria contained in a fluid therein, such as virions in a blood sample. It also provides for an amenable and highly automatable device for massed screenings and the discovery of sequence variants of known virions, previously undetect ed virions, and other genetic material containing moieties. In one embodiment, the present invention utilizes at least one primer for analysis.
Claims (63)
1. A device for storage and analysis of a nucleic acid containing moiety in a biological sample, the device comprising a plurality of layers comprising:
a. a filter layer comprising a dry solid medium comprising a hydrophilic solid matrix; and b. an isolation layer comprising a dry solid medium comprising a neutral solid matrix having sorbed thereto a composition comprising a detergent.
a. a filter layer comprising a dry solid medium comprising a hydrophilic solid matrix; and b. an isolation layer comprising a dry solid medium comprising a neutral solid matrix having sorbed thereto a composition comprising a detergent.
2. The device of claim 1, wherein the detergent comprises SDS.
3. The device of claim 1, wherein the filter layer further comprises an anticoagulant or a liquefying agent.
4. The device of claim 3, wherein the anticoagulant comprises EDTA or Tris-EDTA.
5. The device of claim 3, wherein the liquefying agent comprises thiol.
6. The device of claim 1, wherein the composition of the isolation layer further comprises:
a. a weak base;
b. a chelating agent; and c. optionally, uric acid or a orate salt.
a. a weak base;
b. a chelating agent; and c. optionally, uric acid or a orate salt.
7. The device of claim 6, wherein the weak base comprises Tris.
8. The device of claim 6, wherein the chelating agent comprises EDTA.
9. The device of claim 1, wherein the isolation layer further comprises a primer.
10. The device of claim 1, further comprising c. a cap, wherein the cap comprises:
i. a capping layer comprising a general denaturing agent; and ii. an adhesive edge.
i. a capping layer comprising a general denaturing agent; and ii. an adhesive edge.
11. The device of claim 10, wherein the general denaturing agent comprises SDS.
12. The device of claim 10, wherein the general denaturing agent comprises:
a. a weak base;
b. a chelating agent;
c. an anionic surfactant or detergent; and d. optionally, uric acid or a orate salt.
a. a weak base;
b. a chelating agent;
c. an anionic surfactant or detergent; and d. optionally, uric acid or a orate salt.
13. The device of claim 12, wherein the weak base comprises Tris.
14. The device of claim 12, wherein the chelating agent comprises EDTA.
15. The device of claim 12, wherein the detergent comprises SDS.
16. A device for storage and analysis of a nucleic acid containing moiety in a biological sample, the device comprising a plurality of layers comprising:
a. a collection layer comprising dry solid medium comprising a neutral porous matrix and a dry anticoagulant or liquefying agent;
b. a filter layer comprising a dry solid medium comprising a solid matrix having a particle size limit that precludes eukaryotic cells, but not prokaryotic cells or virions;
c. an isolation layer comprising a dry solid medium comprising a neutral solid matrix having sorbed thereto a composition comprising a detergent.
a. a collection layer comprising dry solid medium comprising a neutral porous matrix and a dry anticoagulant or liquefying agent;
b. a filter layer comprising a dry solid medium comprising a solid matrix having a particle size limit that precludes eukaryotic cells, but not prokaryotic cells or virions;
c. an isolation layer comprising a dry solid medium comprising a neutral solid matrix having sorbed thereto a composition comprising a detergent.
17. The device of claim 16, further comprising:
d. a separator layer between the filter layer and the isolation layer.
d. a separator layer between the filter layer and the isolation layer.
18. The device of claim 16, wherein the detergent comprises SDS.
19. The device of claim 16, wherein the composition of the isolation layer of part b further comprises:
a. a weak base;
b. a chelating agent; and c. optionally, uric acid or a orate salt.
a. a weak base;
b. a chelating agent; and c. optionally, uric acid or a orate salt.
20. The device of claim 19, wherein the weak base comprises Tris.
21. The device of claim 19, wherein the chelating agent comprises EDTA.
22. The device of claim 16, wherein the isolation layer further comprises a primer.
23. The device of claim 16, wherein the filter layer further comprises an anticoagulant or a liquefying agent.
24. The device of claim 23, wherein the anticoagulant comprises EDTA or Tris-EDTA.
25. The device of claim 23, wherein the liquefying agent comprises thiol.
26. The device of claim 16, further comprising:
d. a cap, wherein the cap comprises:
i. a capping layer comprising a general denaturing agent; and ii. an adhesive edge.
d. a cap, wherein the cap comprises:
i. a capping layer comprising a general denaturing agent; and ii. an adhesive edge.
27. The device of claim 26, wherein the general denaturing agent comprises SDS.
28. The device of claim 26, wherein the general denaturing agent comprises:
a. a weak base;
b. a chelating agent;
c. an anionic surfactant or detergent; and d. optionally, uric acid or a orate salt.
a. a weak base;
b. a chelating agent;
c. an anionic surfactant or detergent; and d. optionally, uric acid or a orate salt.
29. The device of claim 28, wherein the weak base comprises Tris.
30. The device of claim 28, wherein the chelating agent comprises EDTA.
31. The device of claim 28, wherein the detergent comprises SDS.
32. A method for storage and analysis of a nucleic acid containing moiety in a biological sample, the device comprising a plurality of layers comprising:
a. applying a biological sample to a filter layer comprising a dry solid medium comprising a hydrophilic solid matrix;
b. filtering the components of the biological sample through the filter layer to an isolation layer comprising a dry solid medium comprising a neutral solid matrix having sorbed thereto a composition comprising a detergent;
c. retaining nucleic acid components in the isolation layer while removing the plurality of non-nucleic acid components;
d. drying the isolation layer;
e. providing a primer; and f. analyzing the nucleic acid components using at least one primer.
a. applying a biological sample to a filter layer comprising a dry solid medium comprising a hydrophilic solid matrix;
b. filtering the components of the biological sample through the filter layer to an isolation layer comprising a dry solid medium comprising a neutral solid matrix having sorbed thereto a composition comprising a detergent;
c. retaining nucleic acid components in the isolation layer while removing the plurality of non-nucleic acid components;
d. drying the isolation layer;
e. providing a primer; and f. analyzing the nucleic acid components using at least one primer.
33. The method of claim 32, wherein the detergent of step b comprises SDS.
34. The method of claim 32, wherein the composition of the isolation layer of step b further comprises:
a. a weak base;
b. a chelating agent; and c. optionally, uric acid or a orate salt.
a. a weak base;
b. a chelating agent; and c. optionally, uric acid or a orate salt.
35. The method of claim 34, wherein the weak base comprises Tris.
36. The method of claim 34, wherein the chelating agent comprises EDTA.
37. The method of claim 32, wherein the filter layer of step a further comprises an anticoagulant or a liquefying agent.
38. The method of claim 37, wherein the anticoagulant comprises EDTA or Tris-EDTA.
39. The method of claim 37, wherein the liquefying agent comprises thiol.
40. The method of claim 32, further comprising a protecting step between step d and step e, wherein the protecting step comprises placing a cap on at least a portion of the device, wherein the cap comprises:
a. a capping layer comprising a general denaturing agent; and b. an adhesive edge.
a. a capping layer comprising a general denaturing agent; and b. an adhesive edge.
41. The method of claim 40, wherein the general denaturing agent comprises SDS.
42. The method of claim 40, wherein the general denaturing agent further comprises:
a. a weak base;
b. a chelating agent;
c. an anionic surfactant or detergent;
d. optionally, uric acid or a orate salt.
a. a weak base;
b. a chelating agent;
c. an anionic surfactant or detergent;
d. optionally, uric acid or a orate salt.
43. The method of claim 42, wherein the weak base comprises Tris.
44. The method of claim 42, wherein the chelating agent comprises EDTA.
45. The method of claim 42, wherein the detergent comprises SDS.
46. The method of claim 32, wherein the isolation layer of step b further comprises a primer.
47. A method for storage and analysis of a nucleic acid containing moiety in a biological sample, the device comprising a plurality of layers comprising:
a. applying a biological sample to a collection layer comprising a neutral porous matrix and a dry anticoagulant or liquefying agent;
b. filtering the remaining components of the biological sample through the collection layer to a filter layer comprising a dry solid medium comprising a solid matrix having a particle size limit that precludes eukaryotic cells, but not prokaryotic cells or virions;
c. filtering the remaining components of the biological sample through the filter layer to a separator layer;
d. filtering the remaining components of the biological sample through the separator layer to an isolation layer comprising a dry solid medium comprising a neutral solid matrix having sorbed thereto a composition comprising a detergent;
e. retaining nucleic acid components in the isolation layer while removing the plurality of non-nucleic acid components;
f. drying the isolation layer;
g. providing at least one primer; and h. analyzing the nucleic acid components using PCR techniques, wherein at least two PCR primers are used.
a. applying a biological sample to a collection layer comprising a neutral porous matrix and a dry anticoagulant or liquefying agent;
b. filtering the remaining components of the biological sample through the collection layer to a filter layer comprising a dry solid medium comprising a solid matrix having a particle size limit that precludes eukaryotic cells, but not prokaryotic cells or virions;
c. filtering the remaining components of the biological sample through the filter layer to a separator layer;
d. filtering the remaining components of the biological sample through the separator layer to an isolation layer comprising a dry solid medium comprising a neutral solid matrix having sorbed thereto a composition comprising a detergent;
e. retaining nucleic acid components in the isolation layer while removing the plurality of non-nucleic acid components;
f. drying the isolation layer;
g. providing at least one primer; and h. analyzing the nucleic acid components using PCR techniques, wherein at least two PCR primers are used.
48. The method of claim 47, wherein the detergent of step d comprises SDS.
49. The method of claim 47, wherein the composition of the isolation layer of step d further comprises:
a. a weak base;
b. a chelating agent; and c. optionally, uric acid or a orate salt.
a. a weak base;
b. a chelating agent; and c. optionally, uric acid or a orate salt.
50. The method of claim 49, wherein the weak base comprises Tris.
51. The method of claim 49, wherein the chelating agent comprises EDTA.
52. The method of claim 47, wherein the isolation layer of step d further comprises a primer.
53. The method of claim 47, wherein the filter layer of step b further comprises an anticoagulant or a liquefying agent.
54. The method of claim 53, wherein the anticoagulant comprises EDTA or Tris-EDTA.
55. The method of claim 53, wherein the liquefying agent comprises thiol.
56. The method of claim 47, further comprising a protecting step between step f and step g, wherein the protecting step comprises placing a cap on at least a portion of the device, wherein the cap comprises:
a. a capping layer comprising a general denaturing agent; and b. an adhesive edge.
a. a capping layer comprising a general denaturing agent; and b. an adhesive edge.
57. The method of claim 56, wherein the general denaturing agent comprises SDS.
58. The method of claim 56, wherein the general denaturing agent comprises:
a. a weak base;
b. a chelating agent;
c. an anionic surfactant or detergent; and d. optionally, uric acid or a orate salt.
a. a weak base;
b. a chelating agent;
c. an anionic surfactant or detergent; and d. optionally, uric acid or a orate salt.
59. The method of claim 58, wherein the weak base comprises Tris.
60. The method of claim 58, wherein the chelating agent comprises EDTA.
61. The method of claim 58, wherein the detergent comprises SDS.
62. A kit for storage and analysis of a nucleic acid containing moiety in a biological sample, wherein the kit comprises:
a. a device for storage and analysis of a nucleic acid containing moiety in a biological sample, the device comprising a plurality of layers comprising:
i. a filter layer comprising a dry solid medium comprising a hydrophilic solid matrix; and ii. an isolation layer comprising a dry solid medium comprising a neutral solid matrix having sorbed thereto a composition comprising a detergent; and b. at least one primer for PCR.
a. a device for storage and analysis of a nucleic acid containing moiety in a biological sample, the device comprising a plurality of layers comprising:
i. a filter layer comprising a dry solid medium comprising a hydrophilic solid matrix; and ii. an isolation layer comprising a dry solid medium comprising a neutral solid matrix having sorbed thereto a composition comprising a detergent; and b. at least one primer for PCR.
63. A kit for storage and analysis of a nucleic acid containing moiety in a biological sample, wherein the kit comprises:
a. the device comprising a plurality of layers comprising:
i. a collection layer comprising dry solid medium comprising a neutral porous matrix and a dry anticoagulant or liquefying agent;
ii. a filter layer comprising a dry solid medium comprising a solid matrix having a particle size limit that precludes eukaryotic cells, but not prokaryotic cells or virions;
iii. an isolation layer comprising a dry solid medium comprising a neutral solid matrix having sorbed thereto a composition comprising a detergent; and b. at least one primer for PCR.
a. the device comprising a plurality of layers comprising:
i. a collection layer comprising dry solid medium comprising a neutral porous matrix and a dry anticoagulant or liquefying agent;
ii. a filter layer comprising a dry solid medium comprising a solid matrix having a particle size limit that precludes eukaryotic cells, but not prokaryotic cells or virions;
iii. an isolation layer comprising a dry solid medium comprising a neutral solid matrix having sorbed thereto a composition comprising a detergent; and b. at least one primer for PCR.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US33600501P | 2001-11-15 | 2001-11-15 | |
| US60/336,005 | 2001-11-15 | ||
| PCT/US2002/036978 WO2003064993A2 (en) | 2001-11-15 | 2002-11-15 | Methods and materials for detecting genetic material |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2477433A1 true CA2477433A1 (en) | 2003-08-07 |
| CA2477433C CA2477433C (en) | 2010-07-06 |
Family
ID=27662930
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2477433A Expired - Fee Related CA2477433C (en) | 2001-11-15 | 2002-11-15 | Methods and materials for detecting genetic material |
Country Status (9)
| Country | Link |
|---|---|
| US (2) | US6869769B2 (en) |
| EP (1) | EP1453484B1 (en) |
| AT (1) | ATE452626T1 (en) |
| AU (1) | AU2002365229A1 (en) |
| BR (1) | BR0206488A (en) |
| CA (1) | CA2477433C (en) |
| DE (1) | DE60234859D1 (en) |
| ES (1) | ES2337665T3 (en) |
| WO (1) | WO2003064993A2 (en) |
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-
2002
- 2002-11-15 BR BRPI0206488-0A patent/BR0206488A/en not_active Application Discontinuation
- 2002-11-15 WO PCT/US2002/036978 patent/WO2003064993A2/en not_active Application Discontinuation
- 2002-11-15 US US10/298,255 patent/US6869769B2/en not_active Expired - Fee Related
- 2002-11-15 ES ES02804828T patent/ES2337665T3/en not_active Expired - Lifetime
- 2002-11-15 DE DE60234859T patent/DE60234859D1/en not_active Expired - Lifetime
- 2002-11-15 AU AU2002365229A patent/AU2002365229A1/en not_active Abandoned
- 2002-11-15 AT AT02804828T patent/ATE452626T1/en not_active IP Right Cessation
- 2002-11-15 CA CA2477433A patent/CA2477433C/en not_active Expired - Fee Related
- 2002-11-15 EP EP02804828A patent/EP1453484B1/en not_active Expired - Lifetime
-
2004
- 2004-12-17 US US11/016,291 patent/US7504363B2/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| EP1453484A2 (en) | 2004-09-08 |
| EP1453484B1 (en) | 2009-12-23 |
| US20050095641A1 (en) | 2005-05-05 |
| US6869769B2 (en) | 2005-03-22 |
| BR0206488A (en) | 2008-08-05 |
| ATE452626T1 (en) | 2010-01-15 |
| WO2003064993A2 (en) | 2003-08-07 |
| AU2002365229A1 (en) | 2003-09-02 |
| DE60234859D1 (en) | 2010-02-04 |
| EP1453484A4 (en) | 2005-03-09 |
| US20030134312A1 (en) | 2003-07-17 |
| CA2477433C (en) | 2010-07-06 |
| ES2337665T3 (en) | 2010-04-28 |
| WO2003064993A3 (en) | 2004-03-11 |
| US7504363B2 (en) | 2009-03-17 |
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